Traditional Chinese medicine composition for treating common cold or influenza and preparation method thereof

Abstract

The invention relates to a traditional Chinese medicine composition for treating common cold or flu and a preparation method thereof, the traditional Chinese medicine composition is prepared from the following components in parts by weight: 398-807 parts of kudzuvine root, 263-524 parts of ephedra, 279-536 parts of cassia twig, 799-1519 parts of lalang grass rhizome, 252-542 parts of fried bitter apricot kernel, 252-542 parts of rhizoma zingiberis and 252-542 parts of liquorice. 276-516 parts by weight of dried ginger and 163-311 parts by weight of licorice root; the traditional Chinese medicine composition is mainly used for relieving exterior syndrome, dispelling cold, promoting salivation, relaxing tendons, ventilating lung and relieving cough.

Description

Technical Field

[0001] The invention belongs to the field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for treating common cold or influenza and a preparation method thereof. Background Art

[0002] The common cold is the most common acute respiratory infectious disease, and clinical manifestations include a series of symptoms such as nasal congestion, runny nose, sneezing, sore throat, chills, fever, and cough. The onset is relatively rapid and occurs in all seasons, but is more common in winter and spring. The common cold is a self-limiting disease, mostly caused by viruses. If combined with bacterial infection, it can lead to worsening of the disease, prolonged illness, various complications, and even endanger the patient's life. Influenza, also known as flu, is an acute respiratory infectious disease caused by influenza virus, with a high incidence rate and strong infectiousness, which can easily cause outbreaks or pandemics. The clinical characteristics are acute onset, manifested as fever, fatigue, and muscle aches all over the body, and may also have symptoms such as nasal congestion, runny nose, and sneezing.

[0003] The Neijing believes that colds are mainly caused by exogenous wind evil. In the theory of traditional Chinese medicine, the six pathogenic evils can all be the cause of colds. Because wind is the "chief of all diseases", it is believed that wind evil is the main cause of colds. It harms people when combined with the seasonal qi, such as cold in winter, heat in spring, summer heat and dampness, dryness in autumn, and dampness in the rainy season. Since the incidence rate is higher in winter and spring, cold and heat are more common, forming wind-cold and wind-heat syndromes. Because wind is light and upward, it often attacks the upper part of the body. Therefore, the Suwen Taiyinyangming Lun says: "Those who are injured by wind will be affected first." The lung is the canopy of all internal organs, and it is located in a high position. It opens to the nose and is responsible for breathing. It is responsible for skin and fur. It is delicate and cannot tolerate evil invasion. Therefore, when external evil invades from the mouth, nose, and skin, the lung defense is affected by evil and is the first to be affected. The location of the cold is in the lung defense, and its basic pathogenesis is that the external pathogen affects the lung defense, resulting in disharmony between the defense and the surface, and the lung fails to clear and descend, especially the disharmony between the defense and the surface. Disharmony between the defense and the surface leads to symptoms such as aversion to cold, fever, headache, body pain, and general discomfort; the lung fails to clear and descend, so symptoms such as nasal congestion, runny nose, sneezing, itchy throat, and sore throat are seen. Wind-cold colds are caused by external pathogens, and the location of the disease is on the surface, which should be a superficial syndrome. The treatment principle should focus on dispelling the external pathogens and clearing the lung qi, while taking into account strengthening the body. According to the characteristics and clinical manifestations of influenza, it is similar to the "epidemic cold", "wind-heat", and "epidemic" recorded in traditional Chinese medicine books. The cause of the disease is mainly seasonal pathogens and epidemic toxins, and it often carries the "combined evil" of the six seasonal evils; the pathogenesis is that the evil attacks the defense and the lung fails to clear and descend. Common clinical syndrome types include wind-heat attacking the defense, wind-cold binding the surface, cold on the surface and heat inside, and damp-heat stagnation.

[0004] Most common colds are caused by rhinoviruses, but can also be caused by parainfluenza viruses, respiratory syncytial viruses, echoviruses or coxsackieviruses. There is currently no specific antiviral drug for the treatment of the common cold, and excessive use of antiviral drugs significantly increases the risk of related adverse reactions. The common cold is treated clinically with a combination of multiple drugs for symptomatic treatment and symptom relief. Commonly used drugs include decongestants, antihistamines, antitussives, expectorants, antipyretics and analgesics. At the same time, pay attention to rest, proper hydration, and indoor air circulation to avoid secondary bacterial infections. The main problems in clinical treatment include: ① Most commonly used clinical drugs have adverse reactions to varying degrees. For example, common adverse reactions of antihistamines include drowsiness and fatigue. Workers in industries such as driving vehicles and ships, climbing operations or operating precision instruments should use them with caution; acetaminophen is the most commonly used antipyretic and analgesic, and excessive use can cause liver damage or even liver necrosis; central antitussives can affect sputum discharge; ibuprofen can increase the severity of infection, etc.; ② The clinical symptoms of colds are diverse, and multiple drugs are often used in combination to relieve symptoms. There are many kinds of cold medicines on the market with different names, but most of them are compound preparations. The combination of multiple drugs often leads to repeated medication and increases the incidence of adverse reactions; ③ Antimicrobial drugs have no clear effect in treating viral colds. According to statistics, about 50% of patients in clinical practice use antimicrobial drugs to treat common colds without the guidance of a doctor. Abuse of antimicrobial drugs can easily induce bacterial resistance and may also cause gastrointestinal reactions.

[0005] Currently, most of the Chinese patent medicines on the market are for treating wind-heat syndrome, and there are only a handful of medicines for treating the common cold (wind-cold syndrome). The medicines recommended in the 2015 edition of the "Guidelines for the Diagnosis and Treatment of Common Colds in Traditional Chinese Medicine" are mainly: Ganmao Qingre Granules, Zhengchaihuyin Granules, Chailian Oral Liquid, Guizhi Granules, etc.

[0006] Therefore, there is an urgent need for a Chinese medicine that can treat the common cold (wind-cold syndrome) or influenza, relieve the symptoms and dispel the cold while preventing the evil spirits from entering the body and turning into heat. At the same time, it can significantly improve the patient's systemic symptoms such as headache and body aches, and has a more comprehensive therapeutic effect and good safety. Summary of the invention

[0007] In order to solve the problems of the prior art, the present invention provides a traditional Chinese medicine composition, which is prepared from seven traditional Chinese medicine raw materials, including kudzu root, ephedra, cassia twig, white grass root, fried bitter almond, dried ginger, and licorice tablets.

[0008] In the formula, kudzu root is pungent and mild, dispelling wind and cold, relieving muscles and raising yang, promoting fluid and relaxing muscles, and is the main drug. Ephedra is pungent and warm, diaphoretic and dispelling exterior symptoms, promoting lung function and relieving cough; cinnamon twig is pungent and warm, diaphoretic and dispelling exterior symptoms, warming and unblocking meridians; the two drugs combined with kudzu root dispelling wind and cold, relieving muscles and relaxing muscles are the assistant drugs. Imperata root is sweet and cold, clearing heat and promoting fluid, clearing heat evil from the internal qi without hurting yin, which can not only help kudzu root to produce fluid, but also prevent excessive pungent and dry power and wind and cold evil from entering the interior and turning into heat; apricot kernel is bitter and warm, dispelling muscles and dispelling cold, relieving cough and relieving asthma, and moistening the intestines and relieving constipation. The lung and large intestine are exterior and interior, which can help hemp and cinnamon increase the function of promoting lung function and relieving cough; dried ginger is pungent and warm, harmonizing the stomach, dispelling cold and unblocking meridians; the three drugs are used together as adjuvant drugs. Licorice is sweet and mild, clearing heat and detoxifying, moistening the lungs and relieving cough, and harmonizing the drugs. The combination of the drugs has the functions of dispelling exterior symptoms and dispelling cold, promoting fluid and relaxing muscles, promoting lung function and relieving cough. The principle of formula composition is in line with the etiology, pathogenesis and formulation principles of wind-cold colds. Compared with existing Chinese patent medicines for wind-cold colds on the market, the treatment characteristics of this prescription are reflected in the fact that while relieving exterior symptoms and dispersing cold, it can also prevent evil qi from entering the interior and turning into heat. At the same time, it can significantly improve the patient's systemic symptoms such as headache and body aches, and the therapeutic effect is more comprehensive.

[0009] The present invention provides a traditional Chinese medicine composition, which is prepared from the following traditional Chinese medicine raw materials in parts by weight:

[0010] 398-807 parts by weight of kudzu root, 263-524 parts by weight of ephedra, 279-536 parts by weight of cassia twig, 799-1519 parts by weight of Imperata root, 252-542 parts by weight of fried bitter almond, 276-516 parts by weight of dried ginger and 163-311 parts by weight of liquorice tablets.

[0011] Preferably, the Chinese medicine composition of the present invention is characterized in that it is made of the following Chinese medicine raw materials in parts by weight:

[0012] 591-646 parts by weight of kudzu root, 402-425 parts by weight of ephedra, 399-437 parts by weight of cassia twig, 1206-1349 parts by weight of Imperata root, 401-418 parts by weight of fried bitter almond, 381-459 parts by weight of dried ginger, and 245-265 parts by weight of liquorice tablets;

[0013] Most preferably, the Chinese medicine composition of the present invention is characterized in that it is made of the following Chinese medicine raw materials in parts by weight: 626 parts of Pueraria root, 418 parts of ephedra, 418 parts of cassia twig, 1253 parts of Imperata root, 418 parts of stir-fried bitter almonds, 418 parts of dried ginger, and 250 parts of licorice tablets.

[0014] The composition of the present invention is in the form of a pharmaceutical preparation, and the pharmaceutical preparation may further include pharmaceutical excipients as required.

[0015] The pharmaceutical preparation of the present invention is selected from the group consisting of tablets, capsules, lozenges, granules, pills, powders, ointments, pills, suspensions, solutions, injections, suppositories, ointments, sprays, drops, pills or patches.

[0016] The present invention further includes a method for preparing the Chinese medicine composition of the present invention, wherein the medicinal materials are weighed, decocted twice with 6-8 times the amount of water, each time for 20-50 minutes, filtered, the filtrate is concentrated under reduced pressure, ethanol is added to make the alcohol content reach 50-70%, allowed to stand, filtered, and the filtrate is decompressed to recover the ethanol, thereby obtaining the composition;

[0017] Alternatively, weigh the medicinal material, add 4-10 times the amount of water to extract for 0.5-10 hours, filter, and concentrate under reduced pressure to a relative density of 1.20-1.35 (70±1° C.).

[0018] Preferably, the preparation method of the traditional Chinese medicine composition of the present invention comprises weighing the medicinal materials, adding 8 times and 6 times the amount of water respectively and boiling them twice, the first time for 30 minutes and the second time for 20 minutes, filtering, and concentrating the filtrate under reduced pressure to a relative density of 1.05-1.10 (70±1°C), adding ethanol to make the alcohol content reach 60%, standing, filtering, and recovering the ethanol under reduced pressure in the filtrate, and concentrating it to a relative density of 1.28-1.35 (60±1°C).

[0019] The present invention further provides application of the traditional Chinese medicine composition in preparing medicine for treating upper respiratory tract infection.

[0020] The present invention further provides the use of the traditional Chinese medicine composition in preparing medicines for common cold and / or influenza.

[0021] Wherein, the common cold is a cold caused by wind and cold.

[0022] The present invention provides a traditional Chinese medicine composition, characterized in that it is made of the following traditional Chinese medicine raw materials in parts by weight:

[0023] 398-807 parts by weight of kudzu root, 263-524 parts by weight of ephedra, 279-536 parts by weight of cassia twig, 799-1519 parts by weight of Imperata root, 252-542 parts by weight of fried bitter almond, 276-516 parts by weight of dried ginger, 163-311 parts by weight of liquorice tablets,

[0024] It is prepared by the following method:

[0025] Weigh the above medicinal materials, add 6-8 times the amount of water and boil twice, each time for 20-50 minutes, filter, decompress the filtrate and concentrate it, add ethanol to make the alcohol content reach 50-70%, let it stand, filter, decompress the filtrate and recover the ethanol, and obtain the product;

[0026] Alternatively, weigh the above medicinal materials, add 4-10 times the amount of water to extract for 0.5-10 hours, filter, and concentrate under reduced pressure to a relative density of 1.20-1.35 (70±1° C.).

[0027] Preferably, the present invention provides a traditional Chinese medicine composition, characterized in that it is made of the following traditional Chinese medicine raw materials in parts by weight: 626 parts of kudzu root, 418 parts of ephedra, 418 parts of cassia twig, 1253 parts of Imperata root, 418 parts of fried bitter almonds, 418 parts of dried ginger, 250 parts of liquorice tablets,

[0028] It is prepared by the following method:

[0029] Weigh the above medicinal materials, add 6-8 times the amount of water and boil twice, each time for 20-50 minutes, filter, decompress the filtrate and concentrate it, add ethanol to make the alcohol content reach 50-70%, let it stand, filter, decompress the filtrate and recover the ethanol, and obtain the product;

[0030] Alternatively, weigh the above medicinal materials, add 4-10 times the amount of water to extract for 0.5-10 hours, filter, and concentrate under reduced pressure to a relative density of 1.20-1.35 (70±1° C.).

[0031] More preferably, the present invention provides a traditional Chinese medicine composition, characterized in that it is made of the following traditional Chinese medicine raw materials in parts by weight: 626 parts of Pueraria root, 418 parts of Ephedra, 418 parts of Cinnamon twig, 1253 parts of Imperata root, 418 parts of fried bitter almonds, 418 parts of dried ginger, 250 parts of liquorice tablets,

[0032] It is prepared by the following method:

[0033] Weigh the above medicinal materials, add 8 times and 6 times the amount of water respectively, and boil twice, the first time for 30 minutes and the second time for 20 minutes, filter, and concentrate the filtrate under reduced pressure to a relative density of 1.05-1.10 (70±1°C), add ethanol to make the alcohol content reach 60%, let stand, filter, and recover the ethanol from the filtrate under reduced pressure and concentrate it to a relative density of 1.28-1.35 (60±1°C).

[0034] The present invention further provides a method for detecting the Chinese medicine composition, wherein the method uses high performance liquid chromatography to detect the content of ephedrine hydrochloride and / or pseudoephedrine hydrochloride therein, and is characterized in that the method comprises the following steps:

[0035] (1) Preparation of test solution: Take a sample and add a diluent to make a solution;

[0036] (2) Preparation of reference solution: Take ephedrine hydrochloride and / or pseudoephedrine hydrochloride standard reference substances, add diluents to prepare respective reference solutions, or mix them to prepare one reference solution;

[0037] (3) Determination method: inject the test solution and the reference solution into a high performance liquid chromatograph to obtain a chromatogram, and calculate the content of ephedrine hydrochloride and / or pseudoephedrine hydrochloride in the test solution based on the chromatogram;

[0038] Wherein, the diluent is methanol, ethanol, water, acetonitrile or a mixture thereof;

[0039] The HPLC conditions are as follows: the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase is composed of phase A and phase B, acetonitrile is mobile phase A, phosphate buffer is mobile phase B, gradient elution, detection wavelength is 200-300nm, column temperature is 25-45°C, flow rate is 0.6-2.0mL / min, injection volume is 5-30μl; the gradient elution program is as follows:

[0040]

[0041] Preferably, in the detection method described in the present invention, the diluent is methanol; the specifications of the chromatographic column are 5 μm, 4.6×250 mm; the flow rate is 1.0 ml / min; the column temperature is 30°C; the detection wavelength is 210 nm; the injection volume is 10 μl; the phosphate buffer preparation process is as follows: weigh 7.80 g of sodium dihydrogen phosphate, add water to dissolve to 1000 ml, adjust the pH to 3.0 with phosphoric acid, pass through a 0.45 μm microporous filter membrane, That is, the solution is obtained; the preparation of the test solution: take about 0.4g of the sample of the Chinese medicine composition described in any one of claims 1 to 3, accurately weigh it, put it in a 10ml volumetric flask, add about 8ml of methanol, ultrasonicate for 20min, add methanol to the scale, shake well, filter, and take the filtrate to obtain; the preparation of the reference solution: take an appropriate amount of ephedrine hydrochloride and / or pseudoephedrine hydrochloride reference substances, accurately weigh them, and add methanol to make a mixed reference solution containing 50μg and 40μg per 1ml respectively.

[0042] The Chinese medicine composition of the present invention has the functions of dispelling cold, promoting body fluid and relaxing muscles, promoting lung function and relieving cough, and is used for the treatment of common cold (wind-cold syndrome).

[0043] The preparation method of the active ingredient of the present invention is obtained through a large number of screenings and optimized through extraction and purification process research. The specific process is as follows:

[0044] (1) Research on extraction process

[0045] The prescription of the present invention is composed of seven herbs such as kudzu root, ephedra, cassia twig, and white imperata root, and mainly contains triterpenes, glycosides, flavonoids, volatile oils, alkaloids and the like. According to the research reports on the effective ingredients of each herbal flavor and their pharmacological effects, and on the basis of keeping the same clinical decoction method, the white imperata root and fried bitter almond are extracted separately or together, and mixed extraction and the like are studied, and the alcohol precipitation process is considered to be added to reduce the dosage. The various processes are screened and evaluated through pharmacodynamic tests, and after the process route is basically determined, the extraction time and the extraction solvent multiple are optimized to obtain a reasonable optimal extraction process.

[0046] 1. Preliminary study on the extraction of Imperata cylindrica and Apricot kernel separately

[0047] Imperata root mainly contains triterpenoid compounds, among which sapogenin is not easily soluble in water. In the study, 70% ethanol was used for extraction alone. After identification by thin layer chromatography, the results showed that the characteristic spots of Imperata root extracted with 70% ethanol were more and more obvious. Therefore, from the perspective of complete component extraction, the extraction process of Imperata root with 70% ethanol is better.

[0048] Bitter almonds mainly contain water-soluble ingredients, amygdalin, which is easily hydrolyzed by amygdalinase or under acidic conditions. The recipe uses fried bitter almonds to eliminate the hydrolysis of amygdalinase, and examines the effects of mixed extraction of fried bitter almonds with other medicinal flavors and single water extraction or appropriate concentration of ethanol extraction on amygdalin. The results of thin layer chromatography identification show that amygdalin cannot be detected when bitter almonds are mixed with other medicinal flavors, while amygdalin spots are obvious when extracted with water or 70% ethanol.

[0049] In addition, the mixed alcohol extraction of Imperata root and bitter almond was investigated. The experimental results showed that the mixed extraction of Imperata root and fried bitter almond with 70% ethanol was simple and reasonable, and ensured the full extraction of relevant components in Imperata root and avoided the hydrolysis and destruction of amygdalin.

[0050] 2. Preliminary study of mixed water extraction

[0051] 2.1 Design of orthogonal experiment

[0052] According to the properties and pharmacological effects of the chemical components of each medicinal flavor in the prescription, three factors, namely, the amount of extraction solvent, extraction time and number of extractions, were investigated. A three-level design experiment was selected for each factor. The total amount of puerarin and ephedrine extracted was used as an indicator for experimental optimization. The levels of each factor are shown in Table 1.

[0053] Table 1 Extraction condition orthogonal factor level table

[0054]

[0055] 2.2 Test methods

[0056] According to the ratio of each medicinal ingredient in the prescription, 100g of each decoction piece was accurately weighed, a total of 9 portions, and the L9(3)4 orthogonal table was used for 9 experiments under parallel operation conditions. The contents of puerarin and ephedrine in the extracts were determined after extraction and converted into the total amount of extract per gram of decoction piece, which is the orthogonal experimental index.

[0057] 2.3 Determination of puerarin and ephedrine content

[0058] Puerarin content determination method: The content of puerarin in the extract was determined by high performance liquid chromatography (Appendix VI D of Part I of the 2000 edition of the Chinese Pharmacopoeia). Chromatographic conditions: octadecylsilane bonded silica gel was used as the filler; methanol-water (25:75) was used as the mobile phase; the detection wavelength was 250nm; the column temperature was 30℃. Preparation of reference solution: Take an appropriate amount of puerarin reference, accurately weigh it, and add methanol to make a solution containing 80μg per 1ml. Preparation of test solution: Accurately weigh about 0.1g of the extract, add 20ml of water for ultrasonic treatment, pass through a D101 macroporous adsorption resin column (8cm long, 0.8cm inner diameter), wash with 10ml of water, discard the water washing liquid, and then slowly elute with methanol, collect the eluate and make it up to 10ml, shake well to obtain the test solution. Determination method: Accurately aspirate 5μl of the reference solution and the test solution, inject them into the liquid chromatograph, and determine them.

[0059] Ephedrine content determination method: The ephedrine content in the extract is determined by high performance liquid chromatography (Appendix VI D of Part I of the 2000 edition of the Chinese Pharmacopoeia). Chromatographic conditions: Use octadecylsilane bonded silica gel as a filler, methanol-B solution (93:7) (B solution preparation: take 0.6ml of phosphoric acid, 0.985g of disodium hydrogen phosphate, dilute to 500ml with water, then add 0.5070g of sodium heptane sulfonate, ultrasonically dissolve, and pass through a 0.45μm microporous filter membrane to obtain), and the detection wavelength is 210nm. Preparation of reference solution: Take an appropriate amount of ephedrine hydrochloride reference, accurately weigh it, and add methanol to make a solution containing 40μg of ephedrine hydrochloride per 1ml. Preparation of test solution: Accurately weigh about 0.1g of the extract, place it in a 10ml volumetric flask, add methanol to ultrasonically dissolve, and dilute to obtain the test solution. Determination method: Accurately pipette 10 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0060] 2.4 Experimental Results

[0061] The results of the determination of the content of puerarin and ephedrine and the total solid extraction rate in the orthogonal test of water extraction process are shown in Table 2:

[0062] Table 2 Water extraction orthogonal test design and test results

[0063]

[0064] 2.5 Comprehensive analysis of test results

[0065] According to the results of intuitive and variance analysis of the test data, and considering the influence of various factors on all indicators, the optimal extraction method combination sequence can be determined as C2B3A2. That is, first add 8 times the amount of water to extract for 1.5 hours, and then add 6 times the amount of water to extract for 1.5 hours. Considering the actual production situation, extraction efficiency and production cost, it is believed that the second extraction only needs 1 hour, so the more reasonable extraction process should be to add 8 times the amount of water to extract for 1.5 hours, and then add 6 times the amount of water to extract for 1 hour.

[0066] 2.6 Study on Concentration Ratio and Alcohol Precipitation Concentration

[0067] In order to investigate the effects of concentration degree and alcohol precipitation concentration on the content of puerarin and ephedrine and the rate of extracting paste, based on the extraction process of first adding 8 times the amount of water to extract for 1.5 hours and then adding 6 times the amount of water to extract for 1 hour, different concentration ratios of 1:1 and 0.8:1 (concentrate: slice volume) and different alcohol precipitation concentrations of 50%, 60%, and 70% were selected for experiments. The results are shown in Table 3:

[0068] Table 3 Effects of concentration degree and alcohol precipitation concentration on puerarin and ephedrine content and paste yield

[0069]

[0070]

[0071] The results show that the concentration ratio has no obvious effect on the content of puerarin and ephedrine and the rate of extracting oil; however, with the increase of ethanol concentration, the content of puerarin and ephedrine and the rate of extracting oil increase, which may be caused by the fact that the higher the ethanol concentration, the larger the volume of alcohol precipitation liquid and the more extracts. Considering the influence of concentration ratio and alcohol precipitation concentration on the content of puerarin and ephedrine and the rate of extracting oil, the process conditions of 1:1 concentration ratio and 60% alcohol precipitation concentration are more suitable.

[0072] 2.7 Pharmacodynamic test screening

[0073] The pharmacodynamic tests were conducted to investigate the effects of factors such as whether Imperata cylindrica and roasted bitter almonds were extracted separately or mixed separately, the extraction time, and the concentration and alcohol precipitation on the efficacy of the product. The 9 sample preparation methods are shown in Table 4:

[0074] Table 4 Pharmacodynamic experimental samples and their preparation process

[0075]

[0076]

[0077] The test results showed that the 3# extract sample had an inhibitory effect on the virus, and all groups of samples had antipyretic effects, among which the 3# extract had a stronger effect and the 2# extract had a more lasting effect. Therefore, the preparation process of the 3# sample was selected for subsequent research.

[0078] 2.8 Optimization of water extraction of mixed medicinal flavors

[0079] Taking into account the results of pharmacodynamic experiments and the preliminary research on the extraction process, a more reasonable extraction process route can be preliminarily screened out, that is, the mixed medicinal flavors are extracted twice with water, and the time should not be too long (the sample extracted in a short time has a stronger pharmacological effect). After the extract is concentrated to a weight ratio of 1:1 with the medicinal pieces, it is precipitated with alcohol to remove impurities, and the ethanol is recovered to obtain the extract.

[0080] 2.9 Design of orthogonal experiment

[0081] On the basis of basically determining the extraction process route, in order to determine more reasonable and stable process parameters, the three parameters of extraction solvent volume (6-10 times), extraction time (20-50min) and whether alcohol precipitation or alcohol precipitation concentration (0, 50%, 60%) were re-examined, and the total amount of puerarin and ephedrine extracted and the total solid yield of the extract were used as indicators for test optimization. The test parameters are shown in Table 5:

[0082] Table 5 Water extraction test parameters

[0083]

[0084] 2.10 Test methods

[0085] According to the ratio of each medicinal flavor in the prescription, 100g of each decoction piece was accurately weighed, a total of 9 portions, and the L9(3)4 orthogonal table was tested 9 times under parallel operation conditions. Extract twice respectively, combine the extracts and concentrate to 1 times the amount of raw medicine, precipitate with alcohol or continue to concentrate to obtain an extract, and let the precipitated sample stand overnight and then recover the ethanol to obtain an extract. The contents of puerarin and ephedrine in the extract were determined respectively, and converted into the total amount of extract per gram of decoction piece according to the extract yield, which is the orthogonal experimental index.

[0086] 2.11 Determination of total solid yield

[0087] Method: Accurately weigh about 5 g of the extract, place it in an evaporating dish that has been dried to constant weight, evaporate it to dryness in a water bath, dry it in an oven at 105°C for 3 hours, move it to a desiccator, leave it for 1 hour, take it out, and quickly and accurately weigh it.

[0088] The total solid yield is calculated as follows:

[0089]

[0090] 2.11 Test results

[0091] The results of the determination of the content of puerarin and ephedrine and the total solid extraction rate in the orthogonal test of water extraction process are shown in Table 6:

[0092] Table 6 Water extraction orthogonal test design and test results

[0093]

[0094] 2.12 Visual analysis of test results of puerarin, ephedrine and total solid extraction rate

[0095] According to the orthogonal test puerarin content and the range R1, the order of factors affecting puerarin extraction rate is (C) alcohol precipitation concentration > (A) solvent amount > (B) extraction time. According to the data analysis of puerarin content, the best extraction test is C3A2B3.

[0096] According to the ephedrine content and the range R2 of the orthogonal test, the order of factors affecting the ephedrine extraction rate is (B) extraction time > (A) solvent amount > (C) alcohol precipitation concentration. According to the data analysis of ephedrine content, the best extraction test is B2A1C1.

[0097] According to the total solid yield of the extract drying and the range R3, the factors affecting the total solid extraction rate are mainly the concentration of alcohol precipitation, followed by the amount of extraction solvent and time. Therefore, the optimal extraction method combination sequence is C3A1B1.

[0098] Taking into account the influence of various factors on all indicators, it can be seen from the above analysis results that the best combination of extraction process conditions is C3B2A2.

[0099] The results of variance analysis are shown in Table 7: The results show that among the factors examined, only factor C (alcohol precipitation concentration) has a significant effect on the paste yield (P<0.05), has a certain effect on the puerarin content (P<0.10), and has no significant effect on the ephedrine content; while the effects of other factors on the various detection indicators have no significant differences.

[0100] Table 7 Results of variance analysis of the significant effects of various factors on puerarin and ephedrine content

[0101]

[0102] F 0.90 (2, 2) = 9.0, F 0.95 (2, 2) = 19.0

[0103] According to the results of intuitive and variance analysis of the test data, and comprehensive consideration of the impact of various factors on all indicators, the optimal extraction method combination sequence can be determined to be C3B2A2. That is, first add 8 times the amount of water to extract for 30 minutes, then add 6 times the amount of water to extract for 20 minutes, concentrate to 1:1, then use 60% ethanol precipitation, and recover the ethanol to obtain the extract.

[0104] According to the ratio of each medicinal flavor in the prescription, each decoction piece was accurately weighed for a total of 200g, a total of 3 portions. The verification was repeated three times according to the C3B2A2 combination conditions, and the results are shown in Table 8 below.

[0105] Table 8 Results of three process verification tests

[0106]

[0107]

[0108] Three repeated experiments showed that under the C3B2A2 combination conditions, the puerarin and ephedrine contents and the total solid yield were ideal, thus determining the optimal production process of water extraction and alcohol precipitation, namely, first adding 8 times the amount of water to extract for 30 minutes, then adding 6 times the amount of water to extract for 20 minutes, concentrating to 1:1, adding ethanol to precipitate to 60%, and recovering the ethanol to obtain the extract.

[0109] 3. Concentration process

[0110] According to the optimized process, the extract was concentrated to 1:1 and then alcohol precipitation was performed. Considering the operability of production and facilitating production control to ensure stable product quality, relative density was determined to be used as the concentration endpoint control index. The relative density of the extract after concentration to 1:1 during seven batches of production was investigated. The results are shown in Table 9:

[0111] Table 9 Relative density of the concentrate after the extract was concentrated to 1:1

[0112]

[0113] The relative density of 7 batches of concentrated solutions at 80°C was in the range of 1.028-1.091. Taking into account the differences in medicinal materials and to ensure product stability, the relative density control range of the extract concentration endpoint was determined to be 1.03-1.08 (80°C).

[0114] The following is an explanation and description of the terms of the present invention:

[0115] Information about prescription drug odors is as follows:

[0116] ① Pueraria root: This product is the dried root of Pueraria lobata (Willd.) Ohwi, a plant of the Leguminosae family. This product should comply with the relevant provisions of the Chinese Pharmacopoeia 2020 edition, Volume 1, page 347, under the item of Pueraria root slices.

[0117] ② Ephedra: This product is the dried herbaceous stem of Ephedra sinica Stapf, a plant of the Ephedraceae family. This product should comply with the relevant provisions of the Chinese Pharmacopoeia 2020 edition, Volume 1, page 333, under the item of Ephedra decoction pieces.

[0118] ③ Cinnamon twig: This product is the dried young branches of Cinnamomum cassia Presl, a plant of the Lauraceae family. This product should comply with the relevant provisions of the 2020 edition of the Chinese Pharmacopoeia, Volume 1, page 288, under the item of Cinnamon Twig Pieces.

[0119] ④ Imperata root: This product is the dried rhizome of Imperata cylindrica Beauv.var.major (Nees) CE Hubb. of the Poaceae family. This product should comply with the Chinese Pharmacopoeia 2020 edition [1] Relevant regulations on page 111 under Imperata cylindrica root decoction pieces.

[0120] ⑤ Fried bitter almonds: This product is the dried mature seeds of the Siberian almond Prunus sibirica L. of the Rosaceae family. This product should comply with the relevant provisions of the Fried Bitter Almond Pieces on page 210 of the 2020 edition of the Chinese Pharmacopoeia.

[0121] ⑥ Dried ginger: This product is the dried rhizome of Zingiber officinale Rosc., a plant of the ginger family. This product should comply with the relevant provisions of the Chinese Pharmacopoeia 2020 edition, Volume 1, page 15, under the item of dried ginger slices.

[0122] ⑦ Licorice Tablets: This product is the dried root and rhizome of the leguminous plant Glycyrrhiza uralensis Fisch. This product should comply with the relevant provisions of the Chinese Pharmacopoeia 2020 edition, Volume 1, page 88, under the item of Licorice Tablets.

[0123] The Chinese medicine composition / Chinese medicine preparation of the present invention is preferably in the form of a unit dose pharmaceutical preparation. The oral preparation is selected from one of capsules, tablets, dripping pills, granules, concentrated pills, oral liquids and mixtures. The injection is selected from one of injection liquids, freeze-dried powder injections and water injections.

[0124] The pharmaceutical composition of the present invention, its oral administration preparation may contain common excipients, such as binders, fillers, diluents, tableting agents, lubricants, disintegrants, colorants, flavoring agents and wetting agents, and the tablets may be coated if necessary.

[0125] Suitable fillers include cellulose, mannitol, lactose and other similar fillers. Suitable disintegrants include starch, polyvinyl pyrrolidone and starch derivatives, such as sodium starch glycolate. Suitable lubricants include, for example, magnesium stearate. Suitable pharmaceutically acceptable wetting agents include sodium lauryl sulfate.

[0126] The pharmaceutical composition of the present invention can be prepared into a solid oral composition by conventional methods such as mixing, filling, tableting, etc. Repeated mixing can distribute the active substance throughout those compositions using a large amount of fillers.

[0127] Oral liquid preparations may be in the form of aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be a dry product that can be reconstituted with water or other suitable carriers before use. Such liquid preparations may contain conventional additives, such as suspending agents, for example sorbitol, syrup, methylcellulose, gelatin, hydroxyethylcellulose, carboxymethylcellulose, aluminum stearate gel or hydrogenated edible fats, emulsifiers, for example lecithin, sorbitan monooleate or gum arabic; non-aqueous carriers (which may include edible oils), for example almond oil, fractionated coconut oil, oily esters such as glycerol esters, propylene glycol or ethanol; preservatives, for example methylparaben or propylparaben or sorbic acid, and, if desired, conventional flavoring or coloring agents.

[0128] For injection, a liquid unit dosage form is prepared containing the active substance of the invention and a sterile carrier. Depending on the carrier and concentration, the compound can be suspended or dissolved. Solutions are usually prepared by dissolving the active substance in a carrier, filtering and sterilizing it before filling it into a suitable vial or ampoule, and then sealing it. Excipients such as a local anesthetic, preservatives and buffers can also be dissolved in this carrier. In order to improve its stability, the composition can be frozen after filling into the vial and the water removed under vacuum.

[0129] The pharmaceutical composition of the present invention can be selectively added with a suitable pharmaceutically acceptable carrier when it is prepared into a medicament. The pharmaceutically acceptable carrier is selected from the group consisting of mannitol, sorbitol, sodium pyrosulfite, sodium bisulfite, sodium thiosulfate, cysteine ​​hydrochloride, thioglycolic acid, methionine, vitamin C, disodium EDTA, sodium calcium EDTA, carbonates, acetates, phosphates of monovalent alkali metals or their aqueous solutions, hydrochloric acid, acetic acid, sulfuric acid, phosphoric acid, amino acids, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivatives, cellulose and its derivatives, alginate, gelatin, polyvinyl pyrrolidone, glycerol, soil temperature 80, agar, calcium carbonate, calcium bicarbonate, surfactants, polyethylene glycol, cyclodextrin, β-cyclodextrin, phospholipid materials, kaolin, talc, calcium stearate, magnesium stearate, etc.

[0130] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0131] Beneficial technical effects of the present invention: 1. According to the functions and indications of the Chinese medicine composition of the present invention, the Chinese medicine composition of the present invention is tested for antipyretic, antibacterial, anti-inflammatory, antitussive and expectorant, sweating and the effect on the immune system. The results show that the Chinese medicine composition of the present invention has a certain antipyretic effect on bacterial infection fever caused by yeast and non-specific infection fever caused by skim milk within the dosage range selected in the experiment; it has a significant inhibitory effect on the occurrence and development of inflammation such as croton oil ear swelling in mice and increased capillary permeability caused by acetic acid; in addition, it can significantly prolong the incubation period of cough caused by ammonia water in mice, reduce the number of coughs, and promote the secretion of tracheal sputum. At the same time, the Chinese medicine composition of the present invention has a significant promoting function on non-specific immunity; it has a very significant inhibitory effect on delayed hypersensitivity reactions in mice; and it has an exciting effect on the sweat glands of rats and can promote the secretion of sweat. In vitro antibacterial tests show that the Chinese medicine composition of the present invention has a significant inhibitory effect on bacteria related to upper respiratory tract infections.

[0132] 2. The Chinese medicine composition of the present invention has significant antipyretic, anti-inflammatory, antibacterial, antitussive, expectorant, sweating and non-specific immune function promotion, and significantly inhibits delayed hypersensitivity reactions. The test results are consistent with its functions and indications, providing a basis for the clinical application of the Chinese medicine composition of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0133] Figure 1 : Normal control group: The epithelium of the sweat gland secretory part is a single layer of cubes, the cell cytoplasm is bright, and there is no vacuoles in the cytoplasm;

[0134] Figure 2: Fenghan Ganmao Granule Group: The epithelium of the sweat gland secretory part is a single-layer cubic shape, the cell cytoplasm is bright, and there are many vacuoles in the cytoplasm, accounting for 2 / 3, and most of the vacuoles are enlarged; Figure 2A The cytoplasm of the sweat glands is multivesicular; Figure 2B It is a large cytoplasmic bulla of the sweat gland;

[0135] Figure 3: The high-dose group of the Chinese medicine composition in Example 1 of the present invention: the epithelium of the sweat gland secretory part is a single-layer cubic shape, the cell cytoplasm is bright, the number of vacuoles in the cytoplasm is large, accounting for 2 / 3, and most of the vacuoles are enlarged; Figure 3A There are numerous vacuoles in the cytoplasm of the sweat glands; Figure 3B The vacuoles in the sweat gland cytoplasm are larger;

[0136] Figure 4: The middle dose group of the Chinese medicine composition in Example 1 of the present invention: the epithelium of the sweat gland secretory part is a single-layer cubic shape, the cell cytoplasm is bright, the number of vacuoles in the cytoplasm is large, accounting for 1 / 2, and a small number of vacuoles are enlarged; Figure 4A 1 / 2 of the sweat gland cytoplasm has vacuoles; Figure 4B Some of the sweat gland cytoplasm have larger vacuoles;

[0137] Figure 5: Low-dose group of the Chinese medicine composition of Example 1 of the present invention: the epithelium of the sweat gland secretory part is a single-layer cubic shape, the cell cytoplasm is bright, the number of vacuoles in the cytoplasm is small, and some vacuoles are enlarged; Figure 5A Sweat glands have few cytoplasmic vacuoles; Figure 5B The sweat gland cytoplasmic vacuoles do not enlarge;

[0138] Figure 6 : Specificity chromatogram (1: methanol, 2: sample, 3: negative sample lacking ephedrine, 4: pseudoephedrine hydrochloride, 5: ephedrine hydrochloride, 6: mixed standard);

[0139] Figure 7 : Standard curve of ephedrine hydrochloride;

[0140] Figure 8 : Standard curve of pseudoephedrine hydrochloride; DETAILED DESCRIPTION

[0141] The present invention is further illustrated by the following examples, but are not intended to limit the present invention.

[0142] Effect experiment part

[0143] Test drug: The fluid extract of the Chinese medicine composition of the present invention was prepared according to Example 1, and each gram of the fluid extract was equivalent to 8.78 g of the raw drug. It was stored in a refrigerator at 4°C and prepared with distilled water to a proper concentration before the test for oral administration to animals.

[0144] The dosage used in the experiment is: the clinical dosage is 180g crude drug / person / day (3g crude drug / kg). Based on this, the dosages used in the experiments of rats and mice are: 72g crude drug / kg (equivalent to 24 times the clinical dosage), 36g crude drug / kg (equivalent to 12 times the clinical dosage), and 18g crude drug / kg (equivalent to 6 times the clinical dosage). The dosages used in the experiments of rabbits are: 36g crude drug / kg, 18g crude drug / kg, and 9g crude drug / kg.

[0145] Positive control drug: Fenghan Ganmao Granules: produced by Bijie Pharmaceutical Factory, Guizhou Province, dosage: 8g powder / kg (20 times the clinical dose). (2) Aspirin: raw material powder, dosage 0.2g / kg.

[0146] Other experimental materials: Croton oil inflammatory agent: prepared at a concentration of 2%; Evans blue solution: prepared at a concentration of 1% with physiological saline; Carrageenan suspension: provided by Liaoning Institute of Pharmacology, prepared at a concentration of 1% with distilled water on the day of use, and shaken for 1 hour to form a suspension. Ammonia water, analytical grade. India ink: produced by Beijing Xizhong Chemical Plant.

[0147] Experimental animals: Km mice, weighing about 20g, both male and female. Produced and provided by the animal room of Tianjin Pharmaceutical Research Institute, certificate number: WJ Jinshi Dongzhi R Zhunzi 001. Wistar rats, weighing about 200g, both male and female. Produced and provided by the animal room of Tianjin Pharmaceutical Research Institute, certificate number: WJ Jinshi Dongzhi R Zhunzi 001. Rabbits: weighing about 2.5kg, half male and half female, purchased from the animal room of our institute. Raised in an observation room with central air conditioning (Tianjin Dongzhi Facility No. 012, in line with barrier level standards), the feed is complete nutrient blocks for rats and special feed for rabbits produced by the China National Institute for the Control of Biological Products, and drinking tap water.

[0148] Effect Example 1 Heat-clearing effect

[0149] 1. Antipyretic effect on yeast-induced fever in rats

[0150] The experiment selected 50 rats and randomly divided them into 5 groups, each group had 10 rats, half of which were male and half were female. The rats were given oral administration for 3 days, once a day, and the control group was given the same volume of distilled water by oral administration. Immediately after the last administration, 10 ml / kg of 5% freshly prepared yeast suspension was subcutaneously injected to induce bacterial infection fever. The rectal temperature was measured before administration and 4, 6, and 8 hours after fever. The average body temperature of each administration group was compared with the average body temperature of the control group, and a statistical T test was performed. The results (see Table 10) showed that the Chinese medicine composition of Example 1 of the present invention had a certain antipyretic effect on fever caused by yeast in rats.

[0151] Table 10 Antipyretic effect of the Chinese medicine composition of the present invention on rats (n=10)

[0152]

[0153] Compared with the control group*p<0.05**p<0.01***p<0.001

[0154] 2. Effects on non-infectious fever in rabbits

[0155] The test selected 30 rabbits (big-eared white), weighing about 2.5kg, and randomly divided them into 5 groups, 6 in each group, both male and female. Continuous gavage administration for 3 days, once a day, the control group was given the same volume of distilled water. After the third administration, subcutaneous injection of skim milk 5ml / immediately caused non-infectious fever. Before administration and 4, 6, and 8 hours after fever, the rectal temperature was measured respectively, and the average rectal temperature at different time points after fever in each administration group was compared with the average value of the control group, and a statistical T test was performed. The results (see Table 11) show that the Chinese medicine composition of the embodiment of the present invention 1 also has a certain antipyretic effect on the non-infectious fever caused by skim milk.

[0156] Table 11 Antipyretic effect of the Chinese medicine composition of Example 1 of the present invention on rabbits (n=6)

[0157]

[0158] Compared with the control group *p<0.05 **p<0.01

[0159] Effect Example 2 Antibacterial Effect

[0160] 1. In vitro antibacterial effect test

[0161] Sources of bacteria used: Staphylococcus aureus: standard strain ATCC-25923, provided by Anhui Provincial Health and Epidemic Prevention Station. Standard strain ATCC-6538 and other clinical isolates were provided by the Affiliated Hospital of Anhui Medical University. Pseudomonas aeruginosa: standard strain ATCC-27853 and other clinical isolates were provided by the Affiliated Hospital of Anhui Medical University. Haemophilus influenzae: standard strain ATCC-58539, purchased from the National Institute for the Control of Pharmaceutical and Biological Products. Klebsiella pneumoniae, Streptococcus B, and Neisseria were all clinical isolates provided by the Affiliated Hospital of Anhui Medical University. Culture medium: glucose broth buffered culture medium, Clark medium, etc.

[0162] The drug solution was diluted to different concentrations (experimental concentration range was 60-0.1 mg / ml) by doubling dilution method, and added to 96-well culture plates at 50ul / well, and then the bacterial solution cultured for 24 hours was diluted by 1:5000, and then added to the culture plate in equal amounts, and cultured at 37°C for 24 hours. The turbidity of the culture solution was observed, and the minimum inhibitory concentration of the drug was determined. The MIC50 was calculated according to the NDST-80 software edited by Sun Ruiyuan. During the experiment, 3 replicate wells were set for each drug concentration, and a drug blank control, a bacterial control and a culture solution control were set at the same time. Each genus of bacteria was tested for 7 strains, and each experiment was repeated 2 times. The results (see Table 12) show that the Chinese medicine composition of Example 1 of the present invention has obvious in vitro antibacterial effects on 6 experimental bacteria, among which the inhibitory effects on Staphylococcus aureus and Group B Streptococcus are particularly obvious.

[0163] Table 12 Inhibitory effect of the Chinese medicine composition of Example 1 of the present invention on bacteria

[0164]

[0165] 2. Protective effect on mice infected with bacteria

[0166] Protective effect on mice infected with Staphylococcus aureus in vivo. Preparation of bacterial solution: Inoculate Staphylococcus aureus into nutrient broth medium, culture at 36℃ for 18h, wash three times with saline by centrifugation, determine the number of viable bacteria in experimental infection by turbidimetry, and dilute with 5% yeast solution to a certain concentration of bacterial suspension for use.

[0167] Km mice were selected, weighing 18-22g, half male and half female, and randomly divided into groups, with 12 animals in each group. The mice were given oral administration once a day according to the dosage shown in Table 13 for 3 consecutive days, and the bacterial control group was given the same volume of distilled water by oral administration. After the third administration, each mouse was intraperitoneally injected with 0.3ml of Staphylococcus aureus (1.5×108 bacteria / ml) (the amount of bacteria that kills 50% of the animals), and then the administration was continued for 7 days. The number of mice surviving and the survival time in each group within 7 days were recorded, and the number of surviving animals in each administration group was compared with that in the bacterial control group, and a ×2 test was performed; the survival time of each group of animals was compared with that in the control group, and a t test was performed. The results (see Table 13) show that the Chinese medicine composition of Example 1 of the present invention has no significant effect on the number of surviving mice and the survival time of mice infected with Staphylococcus aureus in vivo.

[0168] Table 13 Protective effect of the Chinese medicine composition of Example 1 of the present invention on mice infected with Staphylococcus aureus

[0169]

[0170] 3. Protective effect on mice infected with group B streptococcus

[0171] Preparation of bacterial solution: Inoculate group B streptococcus into nutrient broth medium containing bovine serum, culture at 36°C for 18 hours, wash three times with normal saline by centrifugation, determine the number of viable bacteria in the experimental infection by turbidimetry, and dilute with 5% yeast solution to a certain concentration of bacterial suspension for later use.

[0172] Km mice were selected, weighing 18-22g, half male and half female, and randomly divided into groups, with 12 animals in each group. The dosage shown in Table 14 was given by gavage once a day for 3 consecutive days, and the bacterial control group was given the same volume of distilled water by gavage. After the third administration, each mouse was intraperitoneally injected with 0.5ml of group B streptococcus (6×108 bacteria / ml), and then the administration was continued for 7 days. The number of mice surviving and the survival time in each group within 7 days were recorded, and the number of surviving animals in each administration group was compared with the number of surviving animals in the control group, and a ×2 test was performed, and a t test was performed on the survival time. The results (see Table 14) show that the Chinese medicine composition of Example 1 of the present invention has no significant effect on the number of mice surviving and the survival time of mice infected with group B streptococcus in vivo.

[0173] Table 14 Protective effect of the Chinese medicine composition of Example 1 of the present invention on mice infected with Group B Streptococcus

[0174]

[0175] Effect Example 3 Anti-inflammatory Test

[0176] 1. Effect on croton oil-induced ear swelling in mice

[0177] The experiment selected 50 male Km mice, randomly divided into groups, 10 mice in each group. The mice were given oral administration once a day according to the dosage shown in Table 15 for 3 consecutive days, and the control group was given the same volume of distilled water. After the last administration, 0.05 ml of 2% croton oil was dripped into the right ear shell of each mouse to cause inflammation. The mice were killed by dislocating the neck 2 hours after the inflammation, the left and right ears were cut off, and the ear pieces at the same part were punched with a 9mm diameter puncher, weighed, and the difference in weight between the two ears was used as the degree of mouse ear swelling. The average value was calculated, and each drug group was compared with the control group, and a statistical t test was performed. The results (see Table 15) show that each dosage group of the Chinese medicine composition in Example 1 of the present invention has a very significant inhibitory effect on mouse ear swelling.

[0178] Table 15 Effect of the Chinese medicine composition of Example 1 of the present invention on ear swelling in mice

[0179]

[0180] Compared with the control group***p<0.001

[0181] 2. Effects on the Permeability of Mouse Peritoneal Capillaries

[0182] The experiment selected Km mice, male, weighing 18-22g. They were randomly divided into 5 groups, 10 mice in each group, and the drugs were administered orally once a day according to the dosage shown in Table 16. The control group was administered with the same volume of distilled water by oral gavage. The drugs were administered continuously for 7 days. 40 minutes after the last administration, 0.1ml / mouse of Evans blue solution (1%) was injected into the tail vein, and 0.2ml / 10g of glacial acetic acid solution (0.67%) was injected into the peritoneum. After 20 minutes, the animals were killed by dislocating the neck, and the peritoneal cavity was flushed with 5ml of normal saline. The eluate was collected and centrifuged at 2500 rpm for 5 minutes. The supernatant was colorimetrically measured at a wavelength of 580nm using a 751 type spectrophotometer to determine the optical density (OD value). The average OD value of each administration group was compared with the average OD value of the control group, and a t test was performed. The results (see Table 16) show that the high and medium dose groups of the Chinese medicine composition in Example 1 of the present invention can significantly inhibit the increase in capillary permeability of mice caused by acetic acid.

[0183] Table 16 Effect of the Chinese medicine composition of Example 1 of the present invention on capillary permeability in mice

[0184]

[0185] Compared with the control group *p<0.05

[0186] 3. Effect of carrageenan on paw swelling in rats

[0187] This test selected 50 male rats and randomly divided them into 5 groups, 10 rats in each group. The rats were given oral administration once a day according to the dosage shown in Table 17 for 3 consecutive days. 30 minutes after the last administration, 0.05 ml / rat of 1% carrageenan slurry was subcutaneously injected into the right hind foot pad of the rat to induce inflammation. The diameter of 0.5 cm below the ankle joint of the right hind limb of each rat was measured with a projector (magnified 8 times) before and 0.5, 1, 2, and 3 hours after inflammation, and the difference in swelling at different time points before and after inflammation was used as the degree of swelling. The average of each group was compared with the average of the control group, and a statistical t test was performed. The results (see Table 17) showed that the Chinese medicine composition of Example 1 of the present invention did not show obvious anti-inflammatory effect on this model, and only the 36g crude drug / kg dosage group had significant differences at 0.5 and 1 hours after inflammation.

[0188] Table 17 Effect of the Chinese medicine composition of Example 1 of the present invention on carrageenan foot swelling in rats n=10

[0189]

[0190] Compared with the control group *P<0.05 **P<0.01

[0191] Effect Example 4 Antitussive and expectorant effect

[0192] 1. Expectorant effect

[0193] The experiment selected 50 Km mice, weighing about 25±2g, half male and half female, and randomly divided into 5 groups, 10 mice in each group. The mice were given oral administration according to the dosage shown in Table 18, once a day for 3 consecutive days, and the control group was given the same volume of distilled water. 40 minutes after the last administration, 0.4ml / mouse of phenol red solution (2.5%) was injected intraperitoneally, and the mice were killed by air plug injection in the tail vein 30 minutes, fixed in the supine position, the neck was cut open, the trachea was separated, and the trachea from the larynx to the chest was cut off and immersed in 1mlNaHCO3 solution (5%), shaken in a constant temperature air bath at 37℃ for 30 minutes, and the OD value was measured at a wavelength of 545nm. The OD value was compared with the OD value of the standard tube to calculate the phenol red content. The average value of each administration group was compared with the average value of the control group, and a t test was performed. The results (see Table 18) show that each dosage group of the Chinese medicine composition in Example 1 of the present invention has a significant expectorant effect.

[0194] Table 18 Effect of the Chinese medicine composition of Example 1 of the present invention on the expectorant effect of mice

[0195]

[0196] Compared with the control group*p<0.05***p<0.001

[0197] 2. Antitussive effect

[0198] The test selected 50 Km mice, half male and half female, and divided them into groups according to Table 19 and administered by gavage, once a day, for 3 consecutive days. 40 minutes after the last administration, each animal was placed in a spray device, and ammonia water with a concentration of 12.5% ​​was sprayed uniformly for 5 seconds using an oil-free air pump with equal pressure (140mmHg). The animals were taken out and the time was immediately recorded, the cough latency of the mice was recorded, and the number of coughs within 3 minutes was counted. The average value obtained in each group was compared with the average value of the control group, and a t test was performed. The results (see Table 19) show that the Chinese medicine composition of Example 1 of the present invention can significantly prolong the cough latency of mice and reduce the number of coughs.

[0199] Table 19 Antitussive effect of the Chinese medicine composition of Example 1 of the present invention on mice

[0200]

[0201]

[0202] Compared with the control group *p<0.05 **p<0.01

[0203] Effect Example 5 Immune Effect

[0204] 1. Effects on the phagocytic function of the reticuloendothelial system in mice

[0205] In this experiment, 50 mice of Km strain, weighing 18-22g, were randomly divided into groups, 10 mice in each group. The mice were given oral administration according to the dosage shown in Table 20, once a day for 3 consecutive days, and the control group was given the same volume of distilled water. 40 minutes after the last administration, 0.1ml / mouse of 20% India ink was injected into the tail vein, and 20ul of blood was collected from the eye sockets 2 minutes and 10 minutes after the injection, dissolved in 2ml 0.1% Na2CO3 solution, fully shaken, and colorimetrically measured at a wavelength of 650nm using a 42-type biochemical analyzer to measure the optical density OD value. After blood collection, the mice were killed by dislocating the neck, weighed, and the weight of the liver and spleen was measured. The phagocytic index K and phagocytic activity α were calculated according to the following formula. The results (see Table 20) show that the high and medium dose groups of the Chinese medicine composition in Example 1 of the present invention have a significant promoting effect on the phagocytic activity of the mouse reticuloendothelial system.

[0206]

[0207] Table 20 Effect of the Chinese medicine composition of Example 1 of the present invention on the phagocytic function of the reticuloendothelial system in mice

[0208]

[0209] Compared with the control group *p<0.05

[0210] 2. Effects on delayed-type hypersensitivity reactions in mice

[0211] The test selected 50 Km mice, male, weighing about 25g, and randomly divided them. According to the dosage shown in Table 21, the control group was gavaged with the same volume of distilled water. The drug was administered once a day for 9 consecutive days. After the 4th administration, 10% chicken red blood cells 50ul were injected sc into the left posterior foot of each mouse for sensitization, and the drug was continued. On the 5th day of sensitization, 5% chicken red blood cells 50ul were injected sc into the right posterior foot of each mouse for attack. Before the attack and 24 hours after the attack, the width of 0.5cm under the right hind foot joint was measured with a projector (magnification 8 times), and the difference between the two (i.e., the swelling degree) was used as a measure of the delayed hypersensitivity index, and the swelling mean value of each group of administration was compared with the swelling mean value of the control group, and a statistical T test was performed. The results (see Table 21) show that the high and medium dose groups of the Chinese medicine composition of the embodiment of the present invention 1 have a very significant inhibitory effect on the delayed hypersensitivity of mice induced by chicken red blood cells.

[0212] Table 21 Effect of the Chinese medicine composition of Example 1 of the present invention on delayed-type hypersensitivity reaction in mice

[0213]

[0214] ***p<0.001 compared with control group

[0215] Effect Example 6 Sweating Effect

[0216] In this experiment, 50 rats were randomly divided into 5 groups, with 10 rats in each group. The drug was administered by gavage once a day at the dose shown in the above experiment for 3 consecutive days, and the control group was given the same volume of distilled water. After the last administration, the animals were placed in a constant temperature observation room at 22°C. After 2 hours, the skin and subcutaneous tissue of the plantar pad were removed, fixed in formalin solution, dehydrated, embedded, sliced, and HE stained. The changes in the sweat gland epithelial cells of the plantar pads of rats in each group were observed under an optical microscope. For results, see the histopathological photos and the attached drawings in the instruction manual. Figure 1 -Figure 5. Figure 1 Figure 2 is the normal control group: the epithelium of the secretory part of the sweat gland is a single-layer cubic shape, the cell cytoplasm is bright, and no vacuoles are formed in the cytoplasm; Figure 3 is the Fenghan Ganmao Granule Group: the epithelium of the secretory part of the sweat gland is a single-layer cubic shape, the cell cytoplasm is bright, and the number of vacuoles in the cytoplasm is large, accounting for 2 / 3, and most of the vacuoles are enlarged; Figure 2A The cytoplasm of the sweat glands is multivesicular; Figure 2B The sweat gland cytoplasm is a large vesicle; FIG3 is a high-dose group of the Chinese medicine composition of Example 1 of the present invention: the epithelium of the secretory part of the sweat gland is a single-layer cube, the cell cytoplasm is bright, the number of vacuoles in the cytoplasm is large, accounting for 2 / 3, and most of the vacuoles are enlarged; Figure 3A There are numerous vacuoles in the cytoplasm of the sweat glands; Figure 3BThe vacuoles in the cytoplasm of the sweat glands are relatively large; FIG4 is a middle dose group of the Chinese medicine composition in Example 1 of the present invention: the epithelium of the secretory part of the sweat glands is a single-layer cubic shape, the cell cytoplasm is bright, the number of vacuoles in the cytoplasm is relatively large, accounting for 1 / 2, and a small number of vacuoles are enlarged; Figure 4A 1 / 2 of the sweat gland cytoplasm has vacuoles; Figure 4B Some vacuoles in the sweat gland cytoplasm are relatively large; FIG5 is a low-dose group of the Chinese medicine composition in Example 1 of the present invention: the epithelium of the secretory part of the sweat gland is a single-layer cubic shape, the cell cytoplasm is bright, the number of vacuoles in the cytoplasm is relatively small, and some vacuoles are enlarged; Figure 5A Sweat glands have few cytoplasmic vacuoles; Figure 5B The test results show that the high and medium doses of the Chinese medicine composition of Example 1 have a certain excitatory effect on the sweat glands of rats and promote the secretion of sweat.

[0217] Effect Example 7: Experimental study on the efficacy of inhibiting influenza virus in mice

[0218] Experimental materials and reagents: The Chinese medicine composition of the present invention was prepared according to Example 1, and mice were selected from 14-16g Bablc female mice; virus: rat lung adapted influenza virus. A / PR / 8 / 34 (H1N1) virus strain; anhydrous ether, syringe, etc. The therapeutic effects of different concentrations of the Chinese medicine composition of Example 1 of the present invention on the respiratory tract infection of mice with rat lung adapted influenza virus A / PR / 8 / 34 (H1N1) were detected.

[0219] Experimental methods

[0220] Preliminary experiment: Determination of the median lethal dose of influenza virus infection in mice: Influenza virus mouse lung adapted strain A / PR / 8 / 34 (H1N1), passaged through chicken embryos. The HA titer of freshly harvested chicken embryo allantoic fluid is ≥1280, and the bacterial culture is negative. The virus was diluted 10 times in sterile saline, and 8 dilutions were made from 10-1 to 10-8. Five mice were inoculated for each dilution, and each mouse was inoculated with 0.1 ml. Result observation: Death and survival were used as indicators for judgment. Any death within 24 hours was non-specific death and was not counted. Observe every day until the 14th day after infection. Result judgment: Calculate LD50 according to the Reed--Muench formula.

[0221] Formal experiment: Experimental mouse grouping: 14-16g healthy mice were randomly divided into 6 groups with 15 mice in each group: Group 1: 14g Chinese medicine / kg body weight, gavage for 3 days; Group 2: 14g Chinese medicine / kg body weight, gavage for 5 days; Group 3: 3.5g Chinese medicine / kg body weight, gavage for 3 days; Group 4: 3.5g Chinese medicine / kg body weight, gavage for 5 days; Group 5: virus control group; Group 6: normal control group.

[0222] Virus dilution: Select a virus strain whose LD50 has been titrated in mice, dissolve it by soaking in running water, dilute it with sterile saline to contain 15 LD50 per 0.1 ml, and temporarily store it in ice water.

[0223] Virus infection and drug administration: Mice were anesthetized with ether and infected with 15 LD50 influenza viruses through the nasal cavity. Drug administration began 2 hours after virus infection, once in the morning and afternoon each day. The administration route was oral administration, 0.1 ml each time, the dosage was the same as above, and the virus control group and the normal control group were replaced with physiological saline. The drug was administered continuously for 3 or 5 days according to the same method.

[0224] Determination of drug treatment effect: Starting from the day the mice were infected with the virus, the mice were observed for 14 days, the number of deaths was recorded every day, the morbidity, mortality, average life span, drug protection rate and extended life rate of the drug-treated mice were calculated, and compared with the virus control group. After statistical processing, the t value and p value were calculated to judge the treatment effect.

[0225] Experimental results: Mice were infected with influenza virus mouse lung adapted strain A / PR / 8 / 34 (H1N1) through the respiratory tract, and the virus infection dose was 15LD50, with 15 mice in each group. Drug treatment was carried out 2 hours after virus infection. The drug group used two doses of 14g / kg body weight and 3.5g / kg body weight. Each dose was divided into two groups of continuous administration for 3 days and 5 days, and the drug was administered twice a day. (During the continuous administration period, due to drug toxicity, mice in the high-dose group and the 5-day experimental group died) The drug protection rates (%) of the 1st to 4th experimental groups were 36%, 50%, 63%, and 43%, respectively, and the life extension rates (%) were 18%, 4%, 44%, and 20%. The results were statistically processed, and the P value of the third experimental group was <0.01, which was significantly effective compared with the virus control group. The experimental results are shown in Table 22.

[0226] Table 22 In vivo anti-influenza virus efficacy experiment of the Chinese medicine composition of Example 1 of the present invention

[0227]

[0228]

[0229] According to the functions and indications of the Chinese medicine composition of the present invention, the Chinese medicine composition of Example 1 of the present invention was tested for antipyretic, antibacterial, anti-inflammatory, antitussive and expectorant, sweating and the effect on the immune system. The results show that the Chinese medicine composition of Example 1 of the present invention has a certain antipyretic effect on bacterial infection fever caused by yeast and non-specific infection fever caused by skim milk within the dosage range selected in the experiment; it has a significant inhibitory effect on the occurrence and development of inflammation such as croton oil ear swelling in mice and increased capillary permeability caused by acetic acid; in addition, it can significantly prolong the incubation period of cough caused by ammonia water in mice, reduce the number of coughs, and promote the secretion of tracheal sputum. At the same time, the Chinese medicine composition of Example 1 of the present invention has a significant promoting function on non-specific immunity; it has a very significant inhibitory effect on delayed hypersensitivity reactions in mice; and it has an exciting effect on the sweat glands of rats and can promote the secretion of sweat. In vitro antibacterial tests show that the Chinese medicine composition of Example 1 of the present invention has a significant inhibitory effect on bacteria related to upper respiratory tract infections. The Chinese medicine composition of Example 1 of the present invention has significant antipyretic, anti-inflammatory, antibacterial, antitussive, expectorant, sweating and non-specific immune function promotion, and significantly inhibits delayed hypersensitivity reactions. The test results are consistent with its functions and indications, providing a basis for the clinical application of the Chinese medicine composition of the present invention.

[0230] Example 1

[0231] 626g of kudzu root, 418g of ephedra, 418g of cassia twig, 1253g of imperata root, 418g of fried bitter almond, 418g of dried ginger and 250g of liquorice tablets are added with 8 times and 6 times the amount of water respectively, and decocted twice, the first time for 30 minutes and the second time for 20 minutes, filtered, and the filtrate is concentrated under reduced pressure to a relative density of 1.05-1.10 (70±1°C), ethanol is added to make the alcohol content reach 60%, allowed to stand, filtered, and the filtrate is decompressed to recover ethanol and concentrated to a relative density of 1.28-1.35 (60±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0232] Example 2

[0233] 398g of kudzu root, 263g of ephedra, 279g of cassia twig, 799g of imperata root, 252g of fried bitter almond, 276g of dried ginger and 163g of liquorice tablets are added with 8 times and 6 times the amount of water respectively, and boiled twice, the first time for 30 minutes and the second time for 20 minutes, filtered, the filtrate is concentrated under reduced pressure to a relative density of 1.05-1.10 (70±1°C), ethanol is added to make the alcohol content reach 60%, allowed to stand, filtered, the filtrate is decompressed to recover ethanol, and concentrated to a relative density of 1.28-1.35 (60±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0234] Example 3

[0235] 807g of kudzu root, 524g of ephedra, 536g of cassia twig, 1519g of imperata root, 542g of fried bitter almond, 516g of dried ginger and 311g of liquorice tablets are added with 8 times and 6 times the amount of water respectively, and decocted twice, the first time for 30 minutes and the second time for 20 minutes, filtered, and the filtrate is concentrated under reduced pressure to a relative density of 1.05-1.10 (70±1°C), ethanol is added to make the alcohol content reach 60%, allowed to stand, filtered, and the filtrate is concentrated under reduced pressure to recover ethanol and concentrated to a relative density of 1.28-1.35 (60±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0236] Example 4

[0237] 591g of kudzu root, 402g of ephedra, 399g of cassia twig, 1206g of imperata root, 401g of fried bitter almond, 381g of dried ginger and 245g of liquorice tablets are added with 8 times and 6 times the amount of water respectively, and decocted twice, the first time for 30 minutes and the second time for 20 minutes, filtered, the filtrate is concentrated under reduced pressure to a relative density of 1.05-1.10 (70±1°C), ethanol is added to make the alcohol content reach 60%, allowed to stand, filtered, the filtrate is decompressed to recover ethanol, and concentrated to a relative density of 1.28-1.35 (60±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0238] Example 5

[0239] 646g of kudzu root, 425g of ephedra, 437g of cassia twig, 1349g of imperata root, 418g of fried bitter almond, 459g of dried ginger and 265g of liquorice tablets are added with 8 times and 6 times the amount of water respectively, and decocted twice, the first time for 30 minutes and the second time for 20 minutes, filtered, the filtrate is concentrated under reduced pressure to a relative density of 1.05-1.10 (70±1°C), ethanol is added to make the alcohol content reach 60%, allowed to stand, filtered, the filtrate is decompressed to recover ethanol, and concentrated to a relative density of 1.28-1.35 (60±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0240] Example 6

[0241] 626g of kudzu root, 418g of ephedra, 418g of cassia twig, 1253g of imperata root, 418g of fried bitter almond, 418g of dried ginger and 250g of liquorice tablets are added with 8 times and 6 times the amount of water respectively, and decocted twice, the first time for 20 minutes and the second time for 20 minutes, filtered, and the filtrate is concentrated under reduced pressure to a relative density of 1.05-1.10 (70±1°C), ethanol is added to make the alcohol content reach 50%, allowed to stand, filtered, and the filtrate is concentrated under reduced pressure to recover ethanol and concentrated to a relative density of 1.28-1.35 (60±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0242] Example 7

[0243] 626g of kudzu root, 418g of ephedra, 418g of cassia twig, 1253g of imperata root, 418g of fried bitter almond, 418g of dried ginger and 250g of liquorice tablets are added with 8 times and 6 times the amount of water respectively, and decocted twice, the first time for 50 minutes and the second time for 20 minutes, filtered, and the filtrate is concentrated under reduced pressure to a relative density of 1.05-1.10 (70±1°C), ethanol is added to make the alcohol content reach 70%, allowed to stand, filtered, and the filtrate is concentrated under reduced pressure to recover ethanol and concentrated to a relative density of 1.28-1.35 (60±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0244] Example 8

[0245] Weigh 626g of Pueraria root, 418g of Ephedra, 418g of Cassia twig, 1253g of Imperata root, 418g of stir-fried bitter almond, 418g of dried ginger, and 250g of licorice tablets, add 8 times the amount of water, extract for 8 hours, filter, and concentrate under reduced pressure to a relative density of 1.05-1.10 (70±1°C) to obtain an extract, that is, a Chinese medicine composition.

[0246] Preparation of the pharmaceutical preparation of the present invention

[0247] Example 9

[0248] The Chinese medicine extract prepared in Example 1 is used as a Chinese medicine raw material to prepare granules. An appropriate amount of dextrin is added to the Chinese medicine extract obtained in Example 1, and fluidized spray granulation is performed. The granules are dried to prepare 1000 g granules, and the granules are packaged.

[0249] Example 10

[0250] According to the same method as Example 8, add an appropriate amount of dextrin to the Chinese medicine extract obtained in any of Examples 2-7, perform fluidized spray granulation, dry, make 1000g granules, and package.

[0251] Embodiment 11

[0252] The Chinese medicine extract prepared in Example 1 is used as the Chinese medicine raw material to prepare capsules, which are prepared by conventional pharmaceutics techniques. Starch and ethanol can be added as wetting agents, granulated, and encapsulated to obtain the capsules.

[0253] Example 12

[0254] Take 0.5 g of the Chinese medicine composition described in any one of Examples 1-8 and 10.5 g of PEG-6000, mix them evenly, heat to melt, transfer the materials to the drip irrigation of the drip pills, drip the medicine liquid into 6-8° C. liquid paraffin, remove oil, and prepare 400 drip pills.

[0255] Example 13

[0256] Take 0.5 g of the Chinese medicine composition described in any one of Examples 1-8, 4.5 g of glucose, 0.9 g of sodium thiosulfate and 1 ml of distilled water, mix the above components evenly, freeze-dry, and pack into 500 vials to obtain.

[0257] Embodiment 14

[0258] Take 0.5 g of the Chinese medicine composition described in any one of Examples 1 to 8, 5.5 g of mannitol, 0.9 g of calcium sodium edetate and 2 ml of distilled water, mix the above components, freeze-dry, and pack into 300 vials to obtain.

[0259] Embodiment 15

[0260] Take 0.5 g of the Chinese medicine composition described in any one of Examples 1 to 8, 50 g of starch and 50 g of sucrose, mix the above components, granulate them, and press them into tablets to obtain tablets.

[0261] Example 16: Method for detecting the content of ephedrine hydrochloride and pseudoephedrine hydrochloride

[0262] (1) Instruments and Materials

[0263] Instrument: Waters 2695-2489 liquid chromatograph

[0264] Reference substances: ephedrine hydrochloride reference substance (China Food and Drug Inspection Institute, content is 100.0%), pseudoephedrine hydrochloride reference substance (China Food and Drug Inspection Institute, content is 99.8%)

[0265] Test sample: granules prepared according to the method of Example 9 of the present invention; negative sample lacking ephedra;

[0266] Reagents: acetonitrile (chromatographic grade), sodium dihydrogen phosphate (analytical grade), phosphoric acid (analytical grade), methanol (analytical grade), water (Milli-Q water treatment system).

[0267] (2) Chromatographic conditions

[0268] Chromatographic column: Waters XBridge C18 (5 μm, 4.6 × 250 mm);

[0269] Mobile phase: Use acetonitrile as mobile phase A and phosphate buffer as mobile phase B, and perform gradient elution according to Table 23 below; Preparation of phosphate buffer: Weigh 7.80 g of sodium dihydrogen phosphate, add water to dissolve to 1000 ml, adjust the pH to 3.0 with phosphoric acid, and pass through a 0.45 μm microporous filter membrane to obtain.

[0270] Table 23 Gradient elution

[0271]

[0272] Flow rate: 1.0 ml / min; column temperature: 30°C; detection wavelength: 210 nm; injection volume: 10 μl.

[0273] (3) Solution preparation

[0274] Preparation of reference substance solution: Take appropriate amount of ephedrine hydrochloride and pseudoephedrine hydrochloride reference substances, weigh accurately, and add methanol to make mixed reference substance solutions containing 50 μg and 40 μg per 1 ml, respectively.

[0275] Preparation of test solution: Take about 0.4g of the product, weigh accurately, place in a 10ml volumetric flask, add about 8ml of methanol, ultrasonicate for 20min, cool, add methanol to the scale, shake well, filter, and take the filtrate.

[0276] (4) Exclusivity

[0277] Take a negative sample lacking ephedra, prepare a negative test solution according to the test sample preparation method, accurately draw blank solvent (methanol), reference solution, test solution and negative sample solution for testing, record the chromatogram, and there should be no chromatographic peaks in the blank solvent and negative sample at the position corresponding to the reference. The results show that the method used to determine the ephedrine hydrochloride and pseudoephedrine hydrochloride in the granules of the Chinese medicine composition in Example 1 of the present invention has good specificity. The results are shown in the attached figure of the specification Figure 6 .

[0278] (5) System suitability investigation

[0279] The reference solution was sampled for 6 injections in succession according to the above liquid phase method, and the peak areas were recorded. The RSDs of the peak areas of ephedrine hydrochloride and pseudoephedrine hydrochloride were 0.23% and 0.42%, respectively. The results showed that the system had good applicability. See Table 24 and Table 25.

[0280] Table 24 Ephedrine hydrochloride precision test results

[0281]

[0282] Table 25 Pseudoephedrine Hydrochloride Precision Test Results

[0283]

[0284] (6) Linear

[0285] Accurately weigh 50.38 mg of ephedrine hydrochloride reference substance and 40.43 mg of pseudoephedrine hydrochloride reference substance, add methanol and dissolve in a 50 ml volumetric flask, shake well, and obtain a mixed reference substance mother solution with a concentration of 1.0076 mg / ml and 0.8070 mg / ml. Accurately pipette 1 ml, 1 ml, 5 ml, 1 ml, 5 ml, and 5 ml of the mixed reference substance solution into 200 ml, 50 ml, 100 ml, 10 ml, 25 ml, and 10 ml volumetric flasks, dilute with methanol and make up to the scale, shake well, and obtain 6 concentrations of reference substance standard solutions. Inject 10 μl respectively, record the peak area, perform linear regression using the least squares method, and calculate the linear regression equation and correlation coefficient. Using the injection amount of ephedrine hydrochloride (X) as the horizontal coordinate and the peak area (Y) as the vertical coordinate, a linear regression calculation was performed, and the equation of the standard curve was y=2,103.7946x-13,456.8332, r=1.0000. Using the injection amount of pseudoephedrine hydrochloride (X) as the horizontal coordinate and the peak area (Y) as the vertical coordinate, a linear regression calculation was performed, and the equation of the standard curve was y=2,141.5663x-15,177.3802, r=1.0000. The results show that the injection amount of ephedrine hydrochloride in the range of 50-5038ng and the injection amount of pseudoephedrine hydrochloride in the range of 40-4035ng show a good linear relationship. See Tables 26 and 27, and the attached drawings of the specification Figure 7 and Figure 8 .

[0286] Table 26 Ephedrine hydrochloride standard curve determination results

[0287]

[0288] Table 27 Pseudoephedrine Hydrochloride Standard Curve Determination Results

[0289]

[0290] (7) Stability

[0291] Stability of reference solution: Take appropriate amount of ephedrine hydrochloride and pseudoephedrine hydrochloride reference, weigh accurately, add methanol to make mixed reference solution, inject at 0h, 5h, 14h, 21h, 24h, 36h, measure according to the law, record the peak area, calculate RSD as 1.40% and 1.20%. The results show that the reference solution is stable within 36h, see Table 28.

[0292] Table 28 Results of stability test of reference substances

[0293]

[0294] Stability of the test solution: Take the granules of the Chinese medicine composition of the present invention, treat them according to the test preparation method, place them at room temperature, inject them at 0h, 10h, 18h, 24h, and 36h, measure them according to the law, record the peak area, and calculate the peak area RSD of ephedrine hydrochloride and pseudoephedrine hydrochloride, which are 0.42% and 0.58% respectively. The results show that the test solution is stable within 36h. See Table 29.

[0295] Table 29 Results of stability test of test products

[0296]

[0297] (8) Repeatability

[0298] Take 6 samples of the granules of the Chinese medicine composition of the present invention, treat them according to the above test solution preparation method, measure them according to the law, record the peak area, calculate the average content of ephedrine hydrochloride and pseudoephedrine hydrochloride to be 1.2968 mg / g and 0.9609 mg / g respectively, and the RSD values ​​of the content are 0.28% and 0.39%. The results show that the method has good repeatability. See Table 30.

[0299] Table 30 Repeatability test results

[0300]

[0301] (9) Accuracy

[0302] Take a test sample with a known content (the content is calculated according to the average content of the repeatability test), take 6 samples, each about 0.2g, accurately weigh, put in a 10ml volumetric flask, add ephedrine hydrochloride and pseudoephedrine hydrochloride reference substances, prepare according to the test sample preparation method, determine according to the law, and calculate the sample recovery rate. The range of ephedrine hydrochloride recovery rate is 95.88% to 98.71%, and the average recovery rate is 97.55%. The range of pseudoephedrine hydrochloride recovery rate is 95.39% to 96.49%, and the average recovery rate is 95.86%. The results show that the recovery rates of ephedrine hydrochloride and pseudoephedrine hydrochloride are both within the range of 95-102%, indicating that the method has good accuracy. See Table 31 and Table 32.

[0303] Table 31 Ephedrine hydrochloride accuracy test results

[0304]

[0305] Table 32 Pseudoephedrine hydrochloride accuracy test results

[0306]

[0307] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims

1. A Chinese medicine composition, characterized in that: It is made from the following Chinese medicinal raw materials in parts by weight: 398-807 parts by weight of kudzu root, 263-524 parts by weight of ephedra, 279-536 parts by weight of cassia twig, 799-1519 parts by weight of Imperata root, 252-542 parts by weight of fried bitter almond, 276-516 parts by weight of dried ginger and 163-311 parts by weight of liquorice tablets.

2. The Chinese medicine composition according to claim 1, characterized in that: It is made from the following Chinese medicinal raw materials in parts by weight: 591-646 parts by weight of kudzu root, 402-425 parts by weight of ephedra, 399-437 parts by weight of cassia twig, 1206-1349 parts by weight of Imperata root, 401-418 parts by weight of stir-fried bitter almond, 381-459 parts by weight of dried ginger and 245-265 parts by weight of liquorice tablets.

3. The Chinese medicine composition according to claim 2, characterized in that: The medicine is prepared from the following Chinese medicinal raw materials in parts by weight: 626 parts of kudzu root, 418 parts of ephedra, 418 parts of cassia twig, 1253 parts of Imperata root, 418 parts of fried bitter almond, 418 parts of dried ginger and 250 parts of liquorice tablets.

4. The Chinese medicine composition according to any one of claims 1 to 3, characterized in that: The composition is in the form of a pharmaceutical preparation, and the pharmaceutical preparation may further include pharmaceutical excipients as required.

5. The Chinese medicine composition according to claim 4, wherein the pharmaceutical preparation is selected from the group consisting of tablets, capsules, lozenges, granules, pills, powders, ointments, pills, suspensions, solutions, injections, suppositories, ointments, sprays, drops, pills or patches.

6. The method for preparing the Chinese medicine composition according to any one of claims 1 to 3, characterized in that: Weigh the medicinal materials, add 6-8 times the amount of water and boil twice, each time for 20-50 minutes, filter, decompress the filtrate and concentrate it, add ethanol to make the alcohol content reach 50-70%, let it stand, filter, decompress the filtrate and recover the ethanol, and obtain the product; Alternatively, weigh the medicinal material, add 4-10 times the amount of water to extract for 0.5-10 hours, filter, and concentrate under reduced pressure to a relative density of 1.20-1.35 (70±1° C.).

7. The preparation method according to claim 6, characterized in that: Weigh the medicinal materials, add 8 times and 6 times the amount of water respectively, and boil twice, the first time for 30 minutes and the second time for 20 minutes, filter, and concentrate the filtrate under reduced pressure to a relative density of 1.05-1.10 (70±1°C), add ethanol to make the alcohol content reach 60%, let it stand, filter, and recover the ethanol from the filtrate under reduced pressure and concentrate it to a relative density of 1.28-1.35 (60±1°C).

8. Use of the Chinese medicine composition according to any one of claims 1 to 3 in the preparation of a medicine for treating upper respiratory tract infection.

9. Use of the Chinese medicine composition according to any one of claims 1 to 3 in the preparation of medicines for the common cold and / or influenza.

10. The use according to claim 9, wherein the common cold is a cold caused by wind and cold.

11. A Chinese medicine composition, characterized in that: It is made from the following Chinese medicinal raw materials in parts by weight: 398-807 parts by weight of kudzu root, 263-524 parts by weight of ephedra, 279-536 parts by weight of cassia twig, 799-1519 parts by weight of Imperata root, 252-542 parts by weight of fried bitter almond, 276-516 parts by weight of dried ginger, 163-311 parts by weight of liquorice tablets, It is prepared by the following method: Weigh the above medicinal materials, add 6-8 times the amount of water and boil twice, each time for 20-50 minutes, filter, decompress the filtrate and concentrate it, add ethanol to make the alcohol content reach 50-70%, let it stand, filter, decompress the filtrate and recover the ethanol, and obtain the product; Alternatively, weigh the above medicinal materials, add 4-10 times the amount of water to extract for 0.5-10 hours, filter, and concentrate under reduced pressure to a relative density of 1.20-1.35 (70±1° C.).

12. A method for detecting the Chinese medicine composition according to any one of claims 1 to 3, wherein the method adopts high performance liquid chromatography to detect the content of ephedrine hydrochloride and / or pseudoephedrine hydrochloride therein, characterized in that: The method comprises the following steps: (1) Preparation of test solution: Take a sample and add a diluent to make a solution; (2) Preparation of reference solution: Take ephedrine hydrochloride and / or pseudoephedrine hydrochloride standard reference substances, add diluents to prepare respective reference solutions, or mix them to prepare one reference solution; (3) Determination method: inject the test solution and the reference solution into a high performance liquid chromatograph to obtain a chromatogram, and calculate the content of ephedrine hydrochloride and / or pseudoephedrine hydrochloride in the test solution based on the chromatogram; Wherein, the diluent is methanol, ethanol, water, acetonitrile or a mixture thereof; The HPLC conditions are as follows: the chromatographic column is filled with octadecylsilane bonded silica gel, the mobile phase is composed of phase A and phase B, acetonitrile is mobile phase A, phosphate buffer is mobile phase B, gradient elution, detection wavelength is 200-300nm, column temperature is 25-45°C, flow rate is 0.6-2.0mL / min, injection volume is 5-30μl; the gradient elution program is as follows:

13. The method according to claim 12, characterized in that The diluent is methanol; the specifications of the chromatographic column are 5μm, 4.6×250mm; the flow rate is 1.0ml / min; the column temperature is 30℃; the detection wavelength is 210nm; the injection volume is 10μl; the preparation process of the phosphate buffer is as follows: weigh 7.80g of sodium dihydrogen phosphate, add water to dissolve it to 1000ml, adjust the pH to 3.0 with phosphoric acid, and pass it through a 0.45μm microporous filter membrane to obtain; preparation of the test solution: take about 0.4g of the sample of the Chinese medicine composition described in any one of claims 1-3, accurately weigh it, put it in a 10ml volumetric flask, add about 8ml of methanol, ultrasonicate for 20min, add methanol to the scale, shake well, filter, and take the filtrate to obtain; preparation of the reference solution: take an appropriate amount of ephedrine hydrochloride and / or pseudoephedrine hydrochloride reference, accurately weigh it, and add methanol to make a mixed reference solution containing 50μg and 40μg per 1ml respectively.

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